B6.129S2-Sele^tm1Hyn Selp^tm1Hyn/J

Stock number: 008437
Research areas: Cardiovascular Research, Cell Biology Research, Developmental Biology Research, Hematological Research, Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

Mice homozygous for both Sele (selectin, endothelial cell) and Selp (selectin, platelet) mutations are viable and fertile. They are characterized by leukocytosis, spontaneous bacterial infections, dermatitis and defective leukocyte recruitment in many models of inflammation.

Detailed description

Mice homozygous for both the Sele^tm1Hyn Selp^tm1Hyn mutations are viable and fertile. They are characterized by leukocytosis, spontaneous bacterial infections, dermatitis and defective leukocyte recruitment in many models of inflammation.

In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. This is the case for the strain above. It should be noted that the phenotype could vary from that originally described. We will modify the strain description if necessary as published results become available.

Development

The Sele^tm1Hyn mutation was created by replacing the exons encoding the signal peptide, lectin domain, and a portion of the epidermal growth factor domain with a PGK-hygro cassette. To create the Selp^tm1Hyn mutation, a portion of exon 3 encoding 10 amino acids of a signal peptide and 27 amino acids of the lectin domain was deleted by the insertion of PGK-neo cassette. The two mutations were introduced and selected sequentially in 129S2/SvPas-derived D3 embryonic stem (ES) cells. This line has been backcrossed from a mixed C57BL/6 and 129P2 background to C57BL/6 for more than 10 generations.

Allele

Symbol: Selp^tm1Hyn
Name: targeted mutation 1, Richard Hynes
MGI accession ID: MGI:1857245
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Selp
Marker name: selectin, platelet
Chromosome: 1
Strain of origin: 129S2/SvPas
Molecular note: A portion of exon 3 encoding 10 amino acids of a signal peptide and 27 amino acids of the lectin domain was replaced with a PGK-neo cassette. Mice were treated with lipopolysaccharide to augment the normally low levels of selectin in lung and liver tissue. A longer transcript, resulting from aberrant or cryptic splicing involving the neomycin cassette, was detected at low levels in mutant mice by Northern blot analysis. RT-PCR further confirmed the presence of low levels of transcript in homozygous mice. Immunofluorescence analysis of activated platelets and lung sections from homozygous mutant mice showed an absence of encoded protein. These results were further confirmed by flow cytometric analysis of activated platelets and metabolic labeling of lung tissue.

Allele

Symbol: Sele^tm1Hyn
Name: targeted mutation 1, Richard Hynes
MGI accession ID: MGI:1857450
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Sele
Marker name: selectin, endothelial cell
Chromosome: 1
Strain of origin: 129S2/SvPas
Molecular note: A modified D3 ES cell line heterozygous for the Selp^tm1Hyn allele was targeted a second time. The exons encoding the signal peptide, lectin domain, and a portion of the epidermal growth factor domain were replaced by the insertion of a PGK-hygro cassette. Both Northern and RT-PCR analysis revealed an absence of normal message for both genes in extracts from cardiac and pulmonary tissue of homozygous mutant mice. This targeted mutation occurred in cis with the Selp^tm1Hyn mutation. Due to their genomic proximity, the two null alleles should propagate together through the offspring.